一株地膜降解真菌的筛选及其降解性能分析

【目的】分离并鉴定具有聚乙烯材料降解能力的微生物菌株,探究其降解农用地膜的效能,为地膜的微生物降解途径提供支撑。【方法】以线性低密度聚乙烯粉末为唯一碳源的培养物中分离出1株具有降解聚乙烯材料能力的真菌,利用分子生物学方法结合菌株的培养性状对该菌株进行鉴定,通过观察聚乙烯粉末降解情况和测定地膜失重率,结合红外扫描、高分辨场发射扫描电子显微镜分析该菌株对农用地膜的降解效果。【结果】筛选获得1株具有农用地膜降解效果的真菌菌株PT1,经鉴定为桔青霉(Penicillium citrinum),桔青霉PT1菌株能以重均分子量(Mw)2000和400000的聚乙烯粉末作为唯一碳源生长,经红外扫描、电镜观察发现桔青霉PT1可侵蚀传统聚乙烯地膜。桔青霉PT1能快速利用聚酯类生物降解地膜生长,35 d地膜失重率达50%左右。【结论】本文筛选到具有地膜降解特性的桔青霉PT1真菌,丰富了降解聚乙烯材料的微生物类群,同时也为废弃农用地膜的处理提供了环保的处理途径。     

基于啮食泡沫塑料黄粉虫肠道菌群中聚苯乙烯生物降解的探究

【背景】石油基塑料垃圾污染具有高度的化学惰性而不易分解,严重威胁着生态和海洋环境,筛选其高效降解菌并利用生物法治理塑料污染具有重要意义。【目的】聚苯乙烯是其中一种不能降解且用途广泛的塑料。近期有研究报道黄粉虫幼虫可啮食聚苯乙烯泡沫塑料。本文以啮食聚苯乙烯白色泡沫塑料的黄粉虫幼虫为研究材料,以期从肠道菌群中分离聚苯乙烯降解菌。【方法】分别以聚苯乙烯泡沫塑料与麦麸(对照组)为唯一食物来源饲喂黄粉虫幼虫,经过60 d的驯化,采用多种培养基对幼虫肠道提取液及虫体粪便排泄物进行分离、培养;通过形态学特征观察、扫描电镜与16S rRNA基因、ITS序列分析扩增、测定,根据序列信息构建系统进化树确定菌株的分类地位;将真菌菌种分别接种于加有PS膜片与颗粒的液体培养基中振荡培养,对膜片的力学性能变化进行测定,并对聚苯乙烯颗粒失重率进行计算。【结果】从幼虫粪便中分离的2株好氧菌分别被鉴定为Bacillus anthracis Ames PSI-1和Enterobacter cloacae PSI-2,从肠道中分离的真菌被鉴定为Aspergillus niger KHJ-1;电镜检测PS膜片表面出现大量孔洞,并且膜片疏水性减弱而亲水性增强,膜片的断裂伸长率与拉伸强度等力学性能也显著下降;真菌KHJ-1对PS颗粒的60 d降解测定中,发现PS颗粒由5 g原始重量减少了214.8 mg,失重率为4.29%。【结论】黄粉虫肠道中存在可以降解聚苯乙烯塑料的菌株,具有开发为新型降解聚苯乙烯高分子塑料的潜力。     

大麦虫及其肠道微生物对聚乙烯的生物降解

【背景】废旧塑料聚乙烯因具有较高的化学惰性,不易被自然降解而形成长期污染。【目的】探究聚乙烯泡沫塑料对大麦虫生长发育的影响,为大麦虫作为降解聚乙烯泡沫塑料的昆虫推广提供理论依据。【方法】以大麦虫幼虫为研究对象,选用常见的泡沫塑料(聚乙烯),采用4种不同的饲喂方式T1 (麦麸)、T2 (泡沫塑料)、T3 (泡沫塑料+麦麸)、T4 (不饲喂)进行驯化,处理30 d后对大麦虫进行解剖,取肠道内容物于LB培养基中进行富集培养,将富集培养后的菌液加入以聚乙烯(polyethylene,PE)为唯一碳源的LCFBM培养基进行选择性培养,从中筛选分离得到对PE塑料有降解能力的菌株。【结果】取食泡沫塑料30d后,与单一饲喂PE相比,麦麸和PE混合饲喂后大麦虫幼虫的存活率为76%。采用傅里叶变换红外光谱检测发现虫粪组分中主要官能团中峰值明显变化,表明PE长链有断裂现象,并从肠道中分离得到3株可以对PE薄膜边缘造成明显侵蚀的菌株。【结论】大麦虫可取食并消化PE塑料,其肠道内的微生物对PE塑料的降解起到关键作用,研究结果为塑料污染的生物降解提供了科学证据。     

聚乳酸降解菌株筛选鉴定及降解过程优化

【目的】筛选能够降解聚乳酸的微生物, 提高聚乳酸的降解速率并鉴定聚乳酸降解酶种类。【方法】以明胶为唯一碳源, 筛选能够降解聚乳酸的微生物; 通过优化明胶添加浓度、聚乳酸添加量以及金属离子种类提高聚乳酸的降解速率; 通过分析降解过程中代谢产物和酶活力变化明确聚乳酸降解酶类别。【结果】筛选获得一株聚乳酸降解菌株, 鉴定为Lentzea waywayandensis; 经优化培养, 聚乳酸在25 d后失重84.8%; 降解过程中, 检测到乳酸的产生, 体外酶活实验仅检测到蛋白酶活力。【结论】明胶作为唯一碳源适用于聚乳酸降解菌株的筛选; 明胶作为碳源的同时可以作为诱导剂诱导菌株L. waywayandensis降解聚乳酸; 此外, 研究表明蛋白酶在聚乳酸降解过程中发挥重要作用。     

塑料饲养大蜡螟幼虫肠道可培养细菌多样性

【背景】昆虫肠道中存在大量的微生物,是昆虫正常生命活动所必需的。它们能够促进维生素的合成、脂肪和碳水化合物的吸收及利用,同时还可以保护宿主抵御天敌,忍受高温以及促进毒素或异生素的代谢,间接促进资源开发。【目的】研究PE塑料饲喂的大蜡螟幼虫肠道可培养细菌的多样性。【方法】利用16S rRNA基因序列分析技术,结合菌落形态和细胞形态及相关生理生化特征鉴定细菌种类。【结果】从大蜡螟幼虫肠道分离纯化的40株可培养细菌得到16种不同细菌遗传型,分别属于芽孢杆菌科(Bacillaceae)、肠球菌科(Enterococcaceae)、葡萄球菌科(Staphylococcaceae)、莫拉菌科(Moraxellaceae)4个科。其中芽孢杆菌科是肠道可培养细菌的优势细菌种类。结合菌落和细胞形态及生理生化特征,确定肠道可培养细菌为芽孢杆菌属9株、肠球菌属4株、葡萄球菌属2株以及不动杆菌属1株。【结论】通过研究大蜡螟幼虫肠道可培养细菌群落结构组成,可为开展大蜡螟肠道的微生态研究提供相关理论基础。     

一株蒽降解细菌的分离及降解特性研究

【目的】从盐碱土壤中筛选蒽降解菌株并分析其降解特性。【方法】采用极度稀释结果流式细胞检测法筛选分离纯化菌株,通过16S rRNA基因序列分析对菌株进行初步鉴定,采用气质联用仪(GC-MS)分析蒽的降解特性。【结果】从盐碱土壤中筛选出一株高效蒽降解菌株。经过16S rRNA基因序列分析,鉴定该菌株为Demequina salsinemorus BJ1。菌株可以利用蒽作为唯一碳源生长,降解率可达92%。在一定浓度范围内,随着蒽浓度的降低,细菌生长速率变快,降解率升高。添加外加碳源后,细菌生长速率明显变快,而对蒽降解率变低。对萃取中间代谢产物的质谱分析表明,降解蒽的中间代谢产物主要有9,10-anthracenedione (9,10-蒽醌)和Phthalic acid (邻苯二甲酸)等,说明它可能通过邻苯二甲酸途径降解蒽。【结论】筛选得到一株新的耐盐碱蒽降解菌,该菌降解效率高,对修复石油污染的土壤有一定的现实意义。     

高效石油降解菌群的构建及对芳香烃化合物萘的协同降解性能

【背景】石油作为一类混杂有机化合物,一旦产生污染就会对人类和环境造成严重的危害。【目的】从新疆石油污染土壤中分离筛选石油降解菌,为石油污染土壤的生物修复提供数据支持及技术参考。【方法】以石油为唯一碳源,通过富集培养、筛选分离得到123株单菌,根据菌落形态挑选出30个不同形态菌株,通过16S rRNA基因序列确定其种属,构建系统发育树;通过原油降解实验筛选出高效石油降解菌,以芳香烃的标志化合物萘为唯一碳源筛选出高效降解菌株,并分别筛选可降解水杨酸、邻苯二酚的菌株。【结果】分离筛选出5株高效石油降解菌,降解率高于85%;萘、水杨酸和邻苯二酚降解菌株各获得一株,将3种菌株按照1:1:1的接种比例对萘进行降解,萘的降解率从单菌60.74%提升到89.40%,菌株间的分工协作可以提高有机物的降解效率。【结论】筛选得到的菌株丰富了石油降解微生物菌种库,不同微生物菌株之间的分工协作为石油污染物的降解提供了新思路,为进一步研究石油污染治理提供参考。     

可降解吡啶的全食副球菌B21-3的筛选鉴定及降解特性

【背景】吡啶作为一种难降解的有机污染物普遍存在于焦化、炼油、皮革和制药等行业的废水中,并对环境造成危害。【目的】治理废水中残留的有机污染物吡啶,筛选高效降解菌。【方法】采用富集培养和选择培养,以石家庄某污水处理厂的活性污泥为材料进行吡啶降解菌的筛选,通过形态特征、生理生化特性、(G+C)mol%测定及16S rRNA基因序列系统发育分析对筛选到的降解菌进行鉴定,并分析其对吡啶的降解特性。【结果】分离筛选到一株能以吡啶为唯一碳源和氮源生长代谢的降解菌B21-3,经鉴定该菌株为全食副球菌(Paracoccuspantotrophus)。菌株B21-3对吡啶的最适降解温度为32°C,最适降解pH为7.0,吡啶浓度为100mg/L时降解率为48.50%±0.02%;通过逐步提高吡啶初始浓度对菌株进行驯化,驯化后菌株可耐受较高浓度吡啶且吡啶降解率显著增加,吡啶浓度为100 mg/L时驯化后菌株B21-3对吡啶的降解率为90.26%±1.70%。驯化后菌株在含吡啶的无机盐平板上传代培养15代后,对吡啶的降解率为89.39%±2.03%。【结论】菌株B21-3具有较强的吡啶降解能力及降解稳定性,该菌株可作为吡啶污染水体生物修复的潜在资源。     

黄翅大白蚁后肠几丁质降解微生物的分离与鉴定

【目的】从培菌白蚁——黄翅大白蚁后肠微生物菌群中分离能降解几丁质的细菌。【方法】以胶体几丁质为唯一碳源,根据胶体几丁质水解透明圈的大小进行筛选。通过形态学、生理生化以及16SrRNA基因序列分析进行菌株鉴定。【结果】从黄翅大白蚁肠道中筛选到8株能够降解胶体几丁质的细菌,它们分别属于芽孢杆菌属(Bacillus)、短芽孢杆菌属(Brevibacillus)、纤维单胞菌属(Cellulomonas)、指孢囊菌属(Dactylosporangium)、黄杆菌属(Flavobacterium)、类芽孢杆菌属(Paenibacillus)、鞘氨醇单胞菌属(Sphingomonas)和寡养单胞菌属(Stenotrophomonas)。8株菌均具有几丁质酶、β-葡萄糖苷酶和内切葡聚糖酶活性。【结论】从黄翅大白蚁后肠中获得8株能够降解胶体几丁质并具有其他碳水化合物降解酶活性的细菌,这一研究为了解白蚁肠道微生物协助白蚁消化食物机制提供了依据。     

小龙虾肠道产木聚糖酶细菌的分离与鉴定

【背景】小龙虾肠道微生物是小龙虾降解纤维素和半纤维素的主要驱动力。【目的】研究肠道内细菌的相对丰度,为揭示肠道微生物在小龙虾纤维素降解过程中的作用提供理论支撑。【方法】采用纯培养法从小龙虾肠道筛选产木聚糖酶细菌,并且对小龙虾肠道细菌进行16S高通量测序。【结果】形态学和16SrRNA基因分子鉴定表明,筛选到的4株产木聚糖酶细菌均属于芽孢杆菌科芽孢杆菌属;结合进一步的生理生化特征鉴定,结果为:菌株Z-3为枯草芽孢杆菌(Bacillus subtilis),菌株Z-4为贝莱斯芽孢杆菌(Bacillus velezensis),菌株Z-29为蜡状芽孢杆菌(Bacillus cereus),菌株Z-30为高地芽孢杆菌(Bacillus altitudinis);16S rRNA基因高通量测序结果表明:在属水平上,小龙虾肠道细菌主要是Candidatus Bacilloplasma、拟杆菌属、弧菌属、不动杆菌属、Dysgonomonas、Tyzzerella3、气单胞菌属和希瓦氏菌属细菌。【结论】小龙虾肠道内细菌资源丰富,且芽孢杆菌属细菌在木质纤维素降解过程中发挥一定功能。     

六种藓类植物茎的比较解剖学研究

通过对6种藓类植物,即褶叶青藓(Brachythecium salebrosum(Web.et Mohr.)B.S.G.)、湿地匐灯藓(Plagiomnium acutum(Lindb.)Kop.)、侧枝匐灯藓(Plagiomnium maximoviczii(Lindb.)Kop.)、大凤尾藓(Fissidensnobilis Griff.)、大羽藓(Thuidium cymbifolium(Doz.et Molk.)B.S.G.)和大灰藓(Hypnum plumaeforme Wils.)嫩茎和老茎的石蜡切片和显微观察发现,同一藓类植株的嫩茎和老茎,茎结构稳定,不同种藓类植物茎横切面具有不同特征.植物体茎横切面形状、表层细胞的层数、细胞大小和细胞壁厚薄、皮层细胞大小和形状、中轴的有无以及比例等特征可以作为藓类植物的分科分类依据之一.     

Photoregulation of seed germination of wild-type and of an aurea-mutant of tomato

Seed germination of an aurea mutant of tomato ( Lycopersicon esculentum Mill.) is promoted by continuous irradiation with red, far-red or long-wavelength far-red (758 nm) light as well as by cyclic irradiations (5 min red or 5 min far-red/25 min darkness). Far-red light applied immediately after each red does not change the germination behaviour. Seed germination of the isogenic wild-type, cv. UC-105, is promoted by continuous and cyclic red light while it is inhibited by continuous and cyclic far-red light and by continious 758 nm irradiation. Far-red irradiation reverses almost completely the promoting effect of red light. The promoting effect (in the aurea mutant) and the inhibitory effect (in the wild-type) of continuous far-red light do not show photon fluence rate dependency above 20 nmol m⁻² s⁻¹. It is concluded that phytochrome controls tomato seed germination throgh low energy responses in both the wild type and the au mutant. The promoting effect of continuous and cyclic far-red light in the au mutant can be attributed to a greater sensitivity to P_fr.     

Cause of Senescence of Nine Sorts of Flowers

The levels of endogenous phytohormones and respiratory rate in nine sorts of flowers such as Cymbidium faberi Rolfe, Nopalxochia ackermannii Kunth and others were investigated both at full bloom and senescence and meanwhile the effect of exogenous phytohormones on prolonging the blossoms and promoting ethylene production were tested. There is a high content of endogenous ethylene in all the long-lived flowere, about 3–16 folds higer than the short-lived ones. There is a high level of ABA at full blooming flowers of short-lived flowers, in which there is no or only some cytokinins in it, but the ratio of CTK (6BA+zeatin)/ABA is smaller(l.7). The endogenous ABA reached a much higher level at senescence in all nine sorts of flowers, so it is reasonable to consider that it is ABA which plays an important role of regulation in controlling flower's senescence. There is a much higher level of GA3 and zeatin in the long-lived flowers which is not demonstrated in the shortlived ones. The respiratory rate is one of the factors controtling the longevity of flowers, but it does not play a decided role. Application of 6BA and zeatin prolongs distinctly orchid’s longevity, however exogenous IAA through the promotive action on ethylene production, evidently extends the longevity of the flowers of the Nopalxochia ackermannii Kunth.     

龙胆科药用植物化学成分的研究现状

龙胆科植物在我国的分布范围很广,且多数为药用植物,其多数种属的药用植物,至今其化学成分尚未被系统研究。综述了目前龙胆科药用植物的化学成分的研究现状及一般提取方法,对近年来发现的环烯醚萜及裂环烯醚萜类化合物进行了总结,为本科药用植物的更深入研究提供了参考。     

Interconnection of parameters of regulation system of lipid peroxidation and of morphophysiological parameters of mouse liver

A complex analysis of seasonal fluctuations of the mean group parameters of the system of regulation of lipid peroxidation has been performed in liver of Balb/c mice. Association of lipid characteristics and morphophysiological parameters is studied in the Balb/c mouse liver. An inter-connection is revealed between the liver index and the amount of lysoforms of phospholipids, the scale and character of the interconnection differing essentially depending on proportion of phos-phatidylcholine in mouse liver phospholipids.     

The effects of glutamine of the maintenance of embryogenic cultures of Cryptomeria japonica

Summary Embryogenic tissues of sugi (Cryptomeria japonica) were induced on a modified Campbell and Durzan (CD) medium containing 1 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 600 mg l⁻¹ glutamine, and subcultured in the medium of the same composition for over 1 yr. This resulted in a mixed culture of embryogenic and non-embryogenic cells. When embryogenic cells were isolated and cultured independently, their capacity to form embryogenic aggregates was lost. Thus, the non-embryogenic cells present within a mixed culture system were essential to the formation of embryogenic aggregates. When embryogenic tissues were isolated and cultured independently on a high glutamine-containing (2400 mg l⁻¹) medium, dry weights and endogenous levels of glutamine increased, and the tissue could generate a large number of embryogenic aggregates. Amino acid analysis of embryogenic and non-embryogenic cells from the maintenance culture indicated a higher level of glutamine was present in the latter. The high endogenous level of glutamine in the non-embryogenic portion of mixed cell masses may be the supplier of glutamine for maintaining the embryogenic property of the tissues.     

Specificity of action of piperidylcholine derivatives as substrates of cholinesterases of various origin

The review deals with study of enzymologic properties of a novel highly specific acetylcholinesterase substrate, N-(β-acetoxyethyl) piperidinium iodomethylate (“piperidylcholine”), and its 30 derivatives that were tested as effectors of cholinesterases of mammals and various species of Pacific squids. It was proven for the first time that responsible for specificity of action was structure of cyclic ammonium grouping of the alcohol part of molecule of the ester substrate. Analysis of specificity is performed based on enzymatic hydrolysis parameters—activity of catalytic center of cholinesterases and bimolecular constant of the reaction rate that are determined at optimal and low substrate concentrations. Among the specially synthesized group of thioester compounds there is revealed one more highly specific acetylcholinesterase substrate—N-(β-acetoxyethyl) piperidinium.     

Physiological characteristics of various species of strains of carboxydobacteria

Seven strains of aerobic carbon monoxide-oxidizing bacteria (carboxydebacteria) when growing on CO as sole source of carbon and energy had doubling times which ranged from 12–42 h. The activity profiles obtained after discontinuous sucrose density gradient centrifugation indicated that the CO-oxidizing enzymes are soluble and the hydrogenases are membrane-bound in all strains examined. The CO-oxidizing enzymes of Pseudomonas carboxydohydrogena, Pseudomonas carboxydoflava, Comamonas compransoris, and the so far unidentified strains OM2, OM3, and OM4 had a molecular weight of 230,000; that of Achromobacter carboxydus amounted to 170,000. The molecular weights of the CO-oxidizing and H₂-oxidizing enzymes turned out to be identical. The cell sonicates were shown to catalyze the oxidation of both CO and H₂ with methylene blue, thionine, phenazine methosulfate, toluylene blue, dichlorophenolindophenol, cytochrome c or ferricyanide as electron acceptors. Methyl viologen, benzyl viologen, FAD⁺, FMN⁺, and NAD(P)⁺ were not reduced. The spectrum of electron acceptors was identical for all strains tested. Neither free formate, hydrogen nor oxygen gas were involved in the CO-oxidation reaction. Methylene blue was reduced by CO at a 1:1 molar ratio. The results indicate that CO-oxidation by carboxydobacteria is catalyzed by identical or similar enzymes and that the reaction obeys the equation CO+H₂OCO₂+2H⁺+2e^- as previously shown for Pseudomonas carboxydovorans.Dedicated to Otto Kandler remembering almost three decades of enjoyable cooperation     

Modulation of allostery of pyruvate kinase by shifting of an ensemble of microstates

Since the introduction of the concepts of allostery about four decades ago, much advancement has been made in elucidating the structure-function correlation in allostery. However, there are still a number of issues that remain unresolved. In this review we used mammalian pyruvate kinase (PK) as a model system to understand the role of protein dynamics in modulating cooperativity. PK has a triosephosphate isomerase (TIM)(α/β)₈ barrel structural motif. PK is an ideal system to address basic questions regarding regulatory mechanisms about this common (α/β)₈ structural motif. The simplest model accounting for all of the solution thermodynamic and kinetic data on ligand-enzyme interactions involves two conformational states, inactive E^T and active E^R. These conformational states are represented by domain movements. Further studies provide the first evidence for a differential effect of ligand binding on the dynamics of the structural elements, not major secondary structural changes. These data are consistent with our model that allosteric regulation of PK is the consequence of perturbation of the distribution of an ensemble of states in which the inactive E^T and active E^R represent the two extreme end states. Sequence differences and ligands can modulate the distribution of states leading to alterations of functions. The future work includes: defining the network of functionally connected residues; elucidating the chemical principles governing the sequence differences which affect functions; and probing the nature of mutations on the stability of the secondary structural elements, which in turn modulate allostery.